Transduction of anti-cell death protein FNK suppresses graft degeneration after autologous cylindrical osteochondral transplantation.
This study shows that artificial super antiapoptotic FNK protein fused with a protein transduction domain (PTD-FNK) maintains the quality of osteochondral transplant by preventing chondrocyte death. Cylindrical osteochondral grafts were obtained from enhanced green fluorescent protein (EGFP)-expressing transgenic rats, in which living chondrocytes express green fluorescence, and submerged into medium containing PTD-FNK, followed by transplantation into cartilage defects of wild-type rats by impact insertion simulating autologous transplantation. The tissues were histologically evaluated by hematoxylin-eosin and Safranin-O staining. At 1 week, chondrocyte alignment was normal in the PTD-FNK treatment group, whereas all grafts without PTD-FNK treatment showed mixed cluster cell distribution. At 4 weeks, all grafts with PTD-FNK treatment showed almost normal matrix, whereas two grafts without PTD-FNK treatment showed fibrocartilage. Notably, all grafts with PTD-FNK retained high intensity of Safranin-O staining, but all grafts without PTD-FNK largely lost Safranin-O staining. PTD-FNK significantly suppressed a decrease in the survival rate and the density of EGFP-positive cells at 1 and 2 weeks, and this tendency continued at 4 weeks. The results of terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP)-nick end-labeling staining showed that PTD-FNK inhibited cell death, indicating that PTD-FNK protects chondrocyte death and suppresses graft degeneration.
Nakachi
Noriki
N
Department of Biochemistry and Cell Biology, Institute of Development and Aging Sciences, Graduate School of Medicine, Nippon Medical School, 1-396 Kosugi-cho, Nakahara-ku, Kawasaki-city, Kanagawa-pref. 211-8533, Japan.
Asoh
Sadamitsu
S
Watanabe
Nobuyoshi
N
Mori
Takashi
T
Matsushita
Takashi
T
Takai
Shinro
S
Ohta
Shigeo
S
D000818:Animals; D017209:Apoptosis; D002357:Cartilage Diseases / Q000473:pathology / Q000628:therapy*; D002358:Cartilage, Articular / Q000473:pathology* / Q000637:transplantation*; D002470:Cell Survival; D019902:Chondrocytes / Q000378:metabolism / Q000473:pathology; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008297:Male; D010619:Phenazines; D017434:Protein Structure, Tertiary; D051381:Rats; D017207:Rats, Sprague-Dawley; D055647:Rats, Transgenic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000494:pharmacology*; D012394:Rosaniline Dyes; D013194:Staining and Labeling; D051020:bcl-X Protein / Q000235:genetics*; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000235:genetics
D010619:Phenazines; D011993:Recombinant Fusion Proteins; D012394:Rosaniline Dyes; D051020:bcl-X Protein; C485184:enhanced green fluorescent protein; D054322:tat Gene Products, Human Immunodeficiency Virus; D049452:Green Fluorescent Proteins; C035906:Fast Green; C009195:safranine T